본 연구에서는 나노섬유를 기반으로 하는 3 차원적인 환경을 2 개의 층으로 만들어 공배양하여 면역세포와 마우스 폐 상피세포에 알레르겐을 처리하는 연구를 하였다. 알레르겐 Der p 의 농도는 수지상세포의 형태를 변형시키지 않는 농도를 결정하였다. 결정한 농도의 Der p 를 마우스 수지상세포와 폐 상피세포인 MLE-12 세포에 처리하였다. 수지상세포와 MLE-12 세포를 세포배양접시와 나노섬유에서 단일 배양하여 Der p 를 처리하면 2 차원과 3 차원이라는 공간적 차이를 가지고 있어 부착한 세포의 형태에 차이가 있었지만 사이토카인을 측정하였을 때, 세포배양접시와 나노섬유의 큰 차이가 나타나지 않았다. 나노섬유를 2 개의 층으로 만들어 공배양 환경을 구축하여 상층에는 MLE-12 세포를 배양하고, 하층에는 수지상세포를 공배양하여 배양액으로 사이토카인을 측정하였다. Der p 처리 시 TNF-α, IL-6 사이토카인은 나노섬유에서 단일 배양 한 배양액과 큰 차이를 보이지 않았지만 IL-23 사이토카인은 공배양한 배양액에서 가장 큰 증가를 보였다.
수지상세포와 MLE-12 세포와 공배양한 배양액의 전체적인 사이토카인을 관찰하기 위해 사이토카인 배열 키트를 진행하였다. Der p 의 유무에 따라 분비되는 사이토카인을 관찰할 수 있었다. 따라서 공배양 환경을 이용하여 알레르겐과 같은 항원의 유무에 따라 수지상세포와 상피세포 간의 상호작용을 생체 외에서 측정, 관찰할 수 있다.
참고문헌
1. Swamydas, M., et al., Isolation of mouse neutrophils. Curr Protoc Immunol., 2015.
110(1): p. 3.20. 1-3.20. 15.
2. Mannino, D.M., et al., Surveillance for asthma—United States, 1960–1995. MMWr CDC Surveill Summ, 1998. 47(1): p. 1-27.
3. Zarogiannis, S., et al., Anti-interleukin-5 therapy and severe asthma. N Engl J Med, 2009. 360(24): p. 2576.
4. Seaton, A., D.J. Godden, and K. Brown, Increase in asthma: a more toxic environment or a more susceptible population? Thorax, 1994. 49(2): p. 171.
5. Cookson, W., The alliance of genes and environment in asthma and allergy. Nature, 1999. 402(6760): p. 5-11.
6. Prescott, S.L., et al., Transplacental priming of the human immune system to environmental allergens: universal skewing of initial T cell responses toward the Th2 cytokine profile. The Journal of Immunology, 1998. 160(10): p. 4730-4737.
7. Halonen, M., et al., Two subphenotypes of childhood asthma that differ in maternal and paternal influences on asthma risk. Am. J. Respir. Crit. Care Med., 1999. 160(2):
p. 564-570.
8. Stein, R.T., et al., Respiratory syncytial virus in early life and risk of wheeze and allergy by age 13 years. The Lancet, 1999. 354(9178): p. 541-545.
9. Venables, K.M. and M. Chan-Yeung, Occupational asthma. The Lancet, 1997.
349(9063): p. 1465-1469.
10. Platts-Mills, T.A., et al., Dust mite allergens and asthma—a worldwide problem.
Journal of Allergy and Clinical Immunology, 1989. 83(2): p. 416-427.
11. Chu, J.K., et al., Epidemiological studies on the acaroid mite. Korean J. Parasitol., 1967. 5(1): p. 69-75.
12. Athanassiou, C.G., et al., Spatiotemporal distribution of insects and mites in horizontally stored wheat. J. Econ. Entomol., 2005. 98(3): p. 1058-1069.
13. Yong, T.-S. and K.-Y. Jeong, Review on ecology of house dust mites in Korea and
suggestion of a standard survey method. Pediatric Allergy and Respiratory Disease, 2011. 21(1): p. 4-16.
14. Bray, L.J., et al., Multi-parametric hydrogels support 3D in vitro bioengineered microenvironment models of tumour angiogenesis. Biomaterials, 2015. 53: p. 609-620.
15. Taubenberger, A.V., et al., 3D extracellular matrix interactions modulate tumour cell growth, invasion and angiogenesis in engineered tumour microenvironments. Acta biomaterialia, 2016. 36: p. 73-85.
16. Bray, L.J., et al., Hydrogel-based in vitro models of tumor angiogenesis, in 3D Cell Culture. 2017, Springer. p. 39-63.
17. Chwalek, K., L.J. Bray, and C. Werner, Tissue-engineered 3D tumor angiogenesis models: potential technologies for anti-cancer drug discovery. Adv. Drug Deliv. Rev., 2014. 79: p. 30-39.
18. Dahlin, R.L., F.K. Kasper, and A.G. Mikos, Polymeric nanofibers in tissue engineering. Tissue Engineering Part B: Reviews, 2011. 17(5): p. 349-364.
19. Stocco, T.D., et al., Nanofibrous scaffolds for biomedical applications. Nanoscale, 2018. 10(26): p. 12228-12255.
20. Prabhakaran, M.P., L. Ghasemi-Mobarakeh, and S. Ramakrishna, Electrospun composite nanofibers for tissue regeneration. J. Nanosci. Nanotechnol., 2011. 11(4): p.
3039-3057.
21. Li, W.J., et al., Biological response of chondrocytes cultured in three‐dimensional nanofibrous poly (ϵ‐caprolactone) scaffolds. Journal of Biomedical Materials Research Part A: An Official Journal of The Society for Biomaterials, The Japanese Society for Biomaterials, and The Australian Society for Biomaterials and the Korean Society for Biomaterials, 2003. 67(4): p. 1105-1114.
22. Liu, H., et al., Electrospinning of nanofibers for tissue engineering applications.
Journal of Nanomaterials, 2013. 2013.
23. Sankaran, K.K., et al., Nanoarchitecture of scaffolds and endothelial cells in engineering small diameter vascular grafts. Biotechnol. J., 2015. 10(1): p. 96-108.
24. Kim, T.-E., et al., Three-dimensional culture and interaction of cancer cells and dendritic cells in an electrospun nano-submicron hybrid fibrous scaffold. International journal of nanomedicine, 2016. 11: p. 823.
25. Li, W.J., et al., Biological response of chondrocytes cultured in three‐dimensional nanofibrous poly (ϵ‐caprolactone) scaffolds. Journal of Biomedical Materials Research Part A: An Official Journal of The Society for Biomaterials, The Japanese Society for Biomaterials, and The Australian Society for Biomaterials and the Korean Society for Biomaterials, 2003. 67(4): p. 1105-1114.
26. Lutz, M.B., et al., An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. Journal of immunological methods, 1999. 223(1): p. 77-92.
27. Song, M.-G., et al., Role of aquaporin 3 in development, subtypes and activation of dendritic cells. Molecular immunology, 2011. 49(1-2): p. 28-37.
28. Pineda-Torra, I., et al., Isolation, culture, and polarization of murine bone marrow-derived and peritoneal macrophages, in Methods in Mouse Atherosclerosis. 2015, Springer. p. 101-109.
29. Swamydas, M., et al., Isolation of mouse neutrophils. Current protocols in immunology, 2015. 110(1): p. 3.20. 1-3.20. 15.